Journal: iScience

Article Title: Combined metabolomic and transcriptomic profiling approaches reveal the cardiac response to high-fat diet

doi: 10.1016/j.isci.2022.104184

Figure Lengend Snippet:

Article Snippet: Plasmid: pCMV Flag ERR1 , Addgene , #10975.

Techniques: Marker, Extraction, Transfection, Lysis, Luciferase, Protease Inhibitor, RNA Sequencing, Recombinant, Plasmid Preparation, Software

Journal: Nature Communications

Article Title: FGF6 and FGF9 regulate UCP1 expression independent of brown adipogenesis

doi: 10.1038/s41467-020-15055-9

Figure Lengend Snippet: a Gene expression in brown preadipocytes treated with vehicle, FGF6, or FGF9 for 24 h. b Schematic presentation of enChIP experiment. c Relative enrichment of FLII and LRFFIP1 proteins in the dCas9-FLAG IP in cells transfected with Ucp1 gRNAs and GFP control. N = 2 per group. d Counts per million for Flii and Lrrfip1 transcripts in brown preadipocytes treated with vehicle or FGF6 for 8 h. N = 3 per group. e FLII protein level in brown preadipocytes treated with vehicle, FGF6, or FGF9 for 48 h. N = 2 per group. Thirty micrograms of total protein was used for WB. f Ucp1 expression in brown preadipocytes transfected with scrambled siRNA or si Lrrfip1 , followed by treatment with vehicle, FGF6, or FGF9 for 24 h. N = 3 per group. g ERRA protein level in brown preadipocytes treated with vehicle, FGF6, or FGF9 for 72 h. N = 2 per group. Thirty micrograms total protein was used for WB. h Chromatin immunoprecipitation of ERRA on the Ucp1 enhancer in brown preadipocytes treated with vehicle, FGF6, or FGF9 for 48 h. i Chromatin immunoprecipitation of ERRA on the Ucp1 enhancer in white preadipocytes treated with vehicle (DMSO) or PGE2 for 24 h. j Ucp1 expression in brown preadipocytes transfected with scrambled siRNA or si Erra , followed by treatment with vehicle, FGF6, or FGF9 for 24 h. N = 3 per group. k Relative abundance of the indicated proteins detected by mass spectrometry in FLAG IP and IgG samples. N = 4–5. l Western blot for ERRA and FLII in FLAG IP and IgG. FGF6 and FGF9 were used at concentrations of 200 and 100 ng/ml, respectively. m Ucp1 expression in brown preadipocytes transfected with Erra , Flii , and Lrrfip1 cDNA. Data are presented as means ± SEM. Two-way ANOVA. **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05. A representative from a total of two to three independent experiments is shown. Source data are provided as a Source Data file.

Article Snippet: For ERRA IP-MS experiment, murine brown preadipocytes were nucleofected with a plasmid overexpressing DDK-tagged ERRA (MR225753, Origene).

Techniques: Gene Expression, Transfection, Control, Expressing, Chromatin Immunoprecipitation, Mass Spectrometry, Western Blot

Journal: Nature Communications

Article Title: FGF6 and FGF9 regulate UCP1 expression independent of brown adipogenesis

doi: 10.1038/s41467-020-15055-9

Figure Lengend Snippet: a Fgf6 expression in SVF and adipocytes from BAT, ingWAT, and pgWAT, b Fgf9 , and c Fgfr3 expression in SVF and adipocytes from BAT and ingWAT of male mice housed at either 5 °C (cold) or 30 °C (TN) for 7 days. N = 4–5 per group. d Ptges expression in BAT and e ingWAT of mice either placed in cages containing running wheel for 11 days (Trained) or housed at 5 °C temperature for 7 days (cold) compared to the control group housed at room temperature (Sedentary). N = 5–6 per group. f Flii and g Erra expression in ingWAT (right) of male C57BL6 mice either placed in cages containing running wheel for 11 days (Trained) or housed at 5 °C temperature for 7 days (cold) compared to the control group housed at room temperature (Sedentary). N = 5–6 per group. h Top: ERRA protein level in ingWAT of male C57BL6 mice housed at either 5 °C (cold) or 30 °C (TN) for 7 days. Down: quantification of ERRA protein relative to β-tubulin. N = 4–5 per group. i Top: FLII protein level in BAT of male C57BL6 mice housed at either 5 °C (cold) or 30 °C (TN) for 7 days. Down: quantification of FLII protein relative to β-tubulin. N = 4–5 per group. Two-sample t test. j Proposed model for mechanism of FGF6/FGF9-mediated Ucp1 expression. Sixty micrograms of total protein was used for WBs in h , i . Data are presented as means ± SEM. Two-way ANOVA in a – c , one-way ANOVA in d – g , and two-sample t test in h , i . **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05. A representative from a total of two to three independent experiments is shown. Source data are provided as a Source Data file.

Article Snippet: For ERRA IP-MS experiment, murine brown preadipocytes were nucleofected with a plasmid overexpressing DDK-tagged ERRA (MR225753, Origene).

Techniques: Expressing, Control